TRANSFORMASI GEN XILOGLUKANASE PADA BEBERAPA JENIS EKSPLAN SENGON (Paraserianthes falcataria L. Nielsen) MELALUI Agrobacterium tumefaciens

Oleh :
Tri Warseno - M0403010 -

ABSTRACT Sengon (Paraserianthes falcataria L. Nielsen) is one of forestry crop which have been developed and recommended for Industrial Timber Estate (HTI). The economic value of this plant such as pulp, raw material of paper industry (pulp and paper), construction, plywood, and also for energy was widely known. To increase the wood quality of sengon in industry especially for the pulp, sengon plantation in which consist higher cellulose were required. Genetic engineering with the DNA technology for the crop improvement in which the times taken where much shorten compare to conventional one. Overexpression of xyloglucanase in poplar (Populus alba) showed the growth enhancement and improved the cellulose depotition at secondary xylem so wood quality yielded good progressively. The aim of this research was to know the result of xyloglucanase gene transformation via Agrobacterium tumefaciens vector of various explants type of sengon. Callus, cotyledons and cotyledonary nodes were taken away from sengon’s seed which germinated on hormon-free MS medium during 10 days used for transformation. A. tumefaciens carrying recombinant plasmid pAaXEG 300 bearing xyloglucanase and NPT II gene encoding kanamycin resistance were used for transformation process. For cocultivation A. tumefaciens with Optical Density (OD600) = 0,4; 0,6 and 0 (as control) value was used. For callus induction 1mg TDZ and 0,25 mg/IAA were added to MS medium respectively. For the selection purpose 300 mg/l kanamycin and 280 mg/l carbenisillin were added to MS medium. The putative transgenic explants were tested on a gene integration test by PCR methode and the results was checked by gel electroforesis. Putative transgenic explants then moved to the somatic embryo induction medium to induce somatic embryo forming. The results showed that the OD600 0,4 value showed much better than OD600= 0,6 for the all of explants used, accordingly callus yielded highest transformation efficiency value ( 40 %). PCR products examined, resulting showed that six of the nine samples tested showed positif, indicating that xyloglucanase gene has successfully integrated on the plant tissue. Key words: xyloglucanase, Paraserianthes falcataria, explant types, transformation